Quality control and single nucleotide resolution analysis of methylated DNA immunoprecipitation products

Anal Biochem. 2010 Dec 1;407(1):141-3. doi: 10.1016/j.ab.2010.07.013. Epub 2010 Jul 23.

Abstract

DNA methylation patterns are altered in many diseases, and their analysis has become of great interest. Methylated DNA immunoprecipitation (MeDIP) is a simple method to enrich the methylated fraction of the genome. However, it has been difficult to assess the quality and the detailed methylation patterns of the immunoprecipitated DNA. Here we present a simple method for the analysis of the immunoprecipitated DNA at single nucleotide resolution by bisulfite treatment and pyrosequencing of genomic regions. The presented method can be used as an initial quality measure prior to genome-wide read-out technologies such as microarrays and second-generation sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry
  • DNA / isolation & purification
  • DNA Methylation*
  • Immunoprecipitation / methods*
  • Immunoprecipitation / standards
  • Nucleotides / analysis
  • Quality Control
  • Sequence Analysis, DNA / methods*
  • Sequence Analysis, DNA / standards
  • Sulfites / chemistry

Substances

  • Nucleotides
  • Sulfites
  • DNA
  • hydrogen sulfite